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DEVELOPMENT OF GENERIC 'ON SITE' MOLECULAR DIAGNOSTICS FOR EU QUARANTINE PESTS AND PATHOGENS'

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Improved diagnostics for quarantine plant pathogens

Transmission of pathogen infections across country borders has raised an alert in the EU. A European consortium addressed such concerns by improving molecular diagnostic tools used to detect quarantine pathogens.

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Infections can have a serious health and economic impact, emphasising the need for taking precautionary measures such as quarantine of plants to be transported across borders. Despite such measures, unless there are sensitive and specific tools that can diagnose the pathogen at early stages, the spreading of infections cannot be ruled out. Seeking to contribute to this, the EU-funded ‘Development of generic 'on site' molecular diagnostics for EU quarantine pests and pathogens’ (PORT CHECK) project developed and evaluated real-time polymerase chain reaction (PCR) assays for a number of key harmful organisms, including the sudden oak death pathogen (Phytophthora ramorum) and the pinewood nematode (Bursaphelenchus xylophilus). The real-time PCR assay is a technique that involves the amplification of a few DNA sequences into millions of copies, thereby allowing sensitive and quantitative detection of pathogens. Project partners optimised the entire process from sample preparation and setting up of real-time PCR detection reactions to interpretation of results. The participation of small and medium-sized enterprises (SMEs) in the project ensured the availability of prototype equipment and kits, reagents and portable real-time PCR. The generated protocols were validated using infected material and were compiled into a booklet that included the methods for detecting 12 different quarantine pathogens and pests. When put to the test across Europe, it was seen that non-specialist users could generate results equivalent to those achieved in the laboratory in less than two hours, and that this could be done remotely from the laboratory. These encouraging results demonstrated that the PORT CHECK tools and procedures could be exploited by EU Member State plant health-competent laboratories and inspection services as well as at points of entry. Adoption of this new technology should improve plant health inspections and reduce risk of the import and export of harmful organisms, thereby preventing any negative social and environmental effects of such transmission.

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